Differential Regulation of Innate Lymphoid Cells in Human and Murine Oral Squamous Cell Carcinoma.

Oral squamous cell carcinomas (OSCC) remain a major healthcare burden in Asian countries. In Pakistan alone, it is the most common cancer in males and second only to breast cancer in females. Alarmingly, treatment options for OSCC remain limited. With this context, investigations made to explore the inflammatory milieu of OSCC become highly relevant, with the hope of practicing immunotherapeutic approaches to address this highly prevalent tumor. We investigated the newly identified innate lymphoid cells (ILCs) and associated cytokines in well-defined human oral squamous cell carcinoma (OSCC) as well as in a 7,12-dimethylbenz[a]anthracene (DMBA)-induced murine model of OSCC using flow cytometry and quantitative real-time polymerase chain reaction (qPCR). We further went on to explore molecular circuitry involved in OSCC by developing a murine model of OSCC and using an α-Thy1 antibody to inhibit ILCs. Amongst the ILCs that we found in human OSCC, ILC3 (23%) was the most abundant, followed by ILC2 (17%) and ILC1 (1%). Mice were divided into four groups: DMBA (n = 33), DMBA+antibody (Ab) (n = 30), acetone (n = 5), and control (n = 5). In murine OSCC tissues, ILC1 and ILC3 were down-infiltrated, while ILC2 remained unchanged compared to controls. Interestingly, compared to the controls (DMBA group), mice treated with the α-Thy1 antibody showed fewer numbers of large tumors, and a larger percentage of these mice were tumor-free at this study's end point. We present novel data on the differential expansion/downsizing of ILCs in OSCC, which provides a pivotal basis to dive deeper into molecular circuitry and the OSCC tumor niche to devise novel diagnostic, therapeutic, and prognostic strategies to prevent/treat oral cancers.

The Analysis of Facio-Dental Proportions to Determine the Width of Maxillary Anterior Teeth: A Clinical Study.

The present study aimed to analyze mid horizontal facial third proportions, those being the interpupillary, inner intercanthal, and bizygomatic distance modified with golden proportion, The Preston proportion, golden percentage and 70% recurring esthetic dental proportion were used for determining maxillary anterior teeth width. A total of 230 participants took part in this study. The front dental and facial photographs along dental stone cast which were converted to three-dimensional (3D) models were used for evaluation. The mid horizontal facial third proportions showed no significant relationship with maxillary anterior teeth width without modification with dental proportions. Whereas, with modification, no statistically significant difference was found between inner-intercanthal distance by golden percentage and width of central incisors. The bizygomatic distance was greater than intercanine distance. While the interpupillary distance by golden proportion was found to be consistent with intercanine distance in female participants. The modified anterior teeth width was significantly different from measured values, when determined by using the three mid facial proportions with Preston and 70% recurring esthetic dental (RED) proportion. Furthermore, the measured width of maxillary anterior teeth showed no difference when plaster dental casts widths were compared with 3D models. The interpupillary, inner-intercanthal, and bizygomatic distance should not be directly used to determine maxillary anterior teeth width. While maxillary anterior teeth width can be determined by modifying the inner inter-canthal distance with golden percentage and interpupillary distance with golden proportion. Moreover, the midfacial third proportions modified with Preston and 70% recurrent esthetic dental proportion were found to be unreliable for the determination of maxillary anterior teeth widths.

Development of a murine model of oral carcinogenesis: an accelerated tool for biomarker and anti-tumour drug discovery.

Oral squamous cell carcinoma (OSCC) is the most common cancer in Pakistani men and the second most common cancer in women. The objective of our study was to devise a novel accelerated murine model of oral carcinogenesis that can be exploited as a tool to investigate the cancer circuitry involved in OSCC and to identify molecules of diagnostic, therapeutic and prognostic significance. A total of 40 healthy male, 6-8 weeks old, 22 ± 2 gram, Naval Medical Research Institute (NMRI) outbred strain mice were recruited in the experiment. NMRI mice are commonly used for animal experiments in various fields of biology and for drug toxicity. Of these, 25 mice underwent the oral carcinogenesis regimen via topical application of 0.5% 9,10-dimethyl-1,2-benzanthracene (DMBA) on the lower left lip for a maximum of 20 weeks and 15 mice were used as controls (without the carcinogenic regimen). Exophytic tissue masses were harvested, fixed in 10% formalin and stained with haematoxylin and eosin (H&E) for microscopic diagnosis. Additionally, the expression levels of CK 5/6, p53 and Ki-67 were investigated using immunohistochemistry. Of the 25 mice which underwent the carcinogenic regimen, 21 developed moderately differentiated squamous cell carcinoma and 1 showed dysplastic features with foci of invasion. Three mice were found dead with lesion(s). CK 5/6 showed strong positivity (100%) and p53 and Ki-67 showed patchy (<30%) strong positivity in OSCC, suggesting the similarity of our model to human OSCC. We present an accelerated, close-to-human carcinogenesis, model of oral carcinogenesis using DMBA in NMRI mice that can be exploited to study the pathogenesis of oral squamous cell carcinoma and subsequently devise immunotherapy or targeted therapy.

An Analysis of Maxillary Anterior Teeth Crown Width-Height Ratios: A Photographic, Three-Dimensional, and Standardized Plaster Model's Study.

OBJECTIVE: To analyze the width and height ratios of maxillary anterior teeth at different crown levels through photographs, 3D, and plaster dental model techniques in a subset of the Pakistani population. Material and Methods. This clinical study consisted of 230 participants. The maxillary impression, standardized photographs, and models were constructed for crown width and height analysis. The SPSS version 25 was used for statistical analysis. Descriptive statistics were carried out for mean, standard deviation, and percentage calculation of teeth width and height, gender, and age of participants. Paired t-test analysis was carried out to compare the dependent variables (teeth size, width, and height ratios) with independent variables (techniques applied, side disparity). A p value of ≤ 0.05 was considered statistically significant. RESULTS: The mean width and height of maxillary anterior teeth obtained through photographs, 3D, and plater models were statistically different. The 3D dental model analysis showed reliable and accurate results. The mean width and height ratio of teeth were different on both sides of the arch. There was a significant difference (p = 0.001) in crown width-height ratios at different crown levels. CONCLUSION: The width and height ratios in the studied population were different at various crown levels. The dimensions of teeth varied from the incisal to the cervical part of the crown. Hence, rather than relying on a single, fixed ratio of 78% to 80% suggested by researchers for anterior teeth, the clinician should adopt different crown width-height ratios to restore teeth with the optimum esthetic outcome.

Efficacy of zinc sulfate on concurrent chemoradiotherapy induced taste alterations in oral cancer patients: A double blind randomized controlled trial.

OBJECTIVES: To observe the efficacy of zinc sulfate on taste alterations in oral cancer patients receiving concurrent chemotherapy with radiotherapy. METHODS: Seventy patients were randomly assigned to both intervention and control group at Oncology Section of Atomic Energy Medical Centre Karachi from September 2017 to March 2018. One group received zinc sulfate capsules (50 mg TDS daily after meals) and the other group received placebo (thrice after meals). Patients were advised to start taking capsules on the first day of their chemoradiation. Both the groups continued the capsules a month after their CCRT ended. RESULTS: Sweet taste was most effected by cancer and its treatment followed by bitter and salty taste. Sour taste was least effected. When both the groups were compared for four tastes for detection threshold, the differences in observation at 3 stages of median IQR were not significant. For recognition threshold between zinc sulfate and placebo, no significant difference was observed in median IQR for salty taste and bitter taste. However, sweet taste (baseline p-value 0.245, end p-value 0.010, follow-up p-value 0.038) was statistically significant at end of CCRT and follow-up stage and sour taste (baseline p-value 0.24, end p-value 0.006, follow-up p-value 0.898) at end of CCRT only. CONCLUSION: Zinc sulfate was not found to be beneficial in preventing chemoradiation induced taste alterations. Taste and smell alterations are common in patients with cancer and do not receive sufficient support to manage taste alterations. This area requires more research to develop a comprehensive understanding of the nature and its management.

Periodontal status of Pakistani orthodontic patients.

The objective of this study was to evaluate and compare the periodontal status of orthodontic patients and non-orthodontic patients, aged 15-28 years, of both genders. The cross-sectional study included 100 orthodontic and 100 non-orthodontic patients evaluated using a Community Periodontal Index for Treatment Need (CPITN) probe on the index teeth. A questionnaire was distributed to the participants to assess and evaluate the use of oral hygiene aids. The data were analyzed using SPSS version 17, and various comparisons were performed using the chi-square test. The study revealed that there was a statistically significant association in CPITN scores between the orthodontic and non-orthodontic patients (p < 0.01). The study showed that patients undergoing orthodontic treatment have increased plaque accumulation and probing depth resulting in periodontal tissue destruction. Proper oral hygiene practices and interdental aids should be employed to control plaque.

Chemokine receptor 7 promotes tumor migration and invasiveness via the RhoA/ROCK pathway in metastatic squamous cell carcinoma of the head and neck.

Metastatic squamous cell carcinoma of the head and neck (SCCHN) has been shown to express chemokine receptor 7 (CCR7), which can activate signaling pathways to promote invasion and survival of SCCHN cells. We hypothesized that the RhoA/Rho-associated kinase (ROCK) pathway is involved in the CCR7-induced invasion and migration of metastatic SCCHN cells. Thus, using migration, matrigel invasion and scrape wound-healing assays, we elucidated the role of RhoA in mediating CCR7-associated cellular mobility. Pull-down assays and western blotting were used to measure RhoA and its downstream expression. Immunohistochemical staining and analysis were useful in identifying the correlation between CCR7 and RhoA expression and clinicopathological factors. The results showed that inhibition of RhoA/ROCK reduced the tumor cell migration and invasiveness induced by CCL19. Activated RhoA, proline-rich tyrosine kinase-2 (Pyk2) and cofilin induced by CCL19 were elevated, and increased RhoA, Pyk2 and cofilin activity was eliminated by CCR7mAb, RhoA/ROCK and Pyk2 inhibitors, indicating involvement of the RhoA/ROCK-Pyk2-cofilin cascade. In summary, CCR7 via RhoA/ROCK-Pyk2 cofilin pathway promotes invasion and migration of metastatic SCCHN cells.

Periodontal status of Pakistani orthodontic patients

The objective of this study was to evaluate and compare the periodontal status of orthodontic patients and non-orthodontic patients, aged 15–28 years, of both genders. The cross-sectional study included 100 orthodontic and 100 non-orthodontic patients evaluated using a Community Periodontal Index for Treatment Need (CPITN) probe on the index teeth. A questionnaire was distributed to the participants to assess and evaluate the use of oral hygiene aids. The data were analyzed using SPSS version 17, and various comparisons were performed using the chi-square test. The study revealed that there was a statistically significant association in CPITN scores between the orthodontic and non-orthodontic patients (p < 0.01). The study showed that patients undergoing orthodontic treatment have increased plaque accumulation and probing depth resulting in periodontal tissue destruction. Proper oral hygiene practices and interdental aids should be employed to control plaque.

CCR7 regulates cell migration and invasion through JAK2/STAT3 in metastatic squamous cell carcinoma of the head and neck.

Squamous cell carcinoma of the head and neck (SCCHN) frequently involves metastasis at diagnosis. Our previous research has demonstrated that CCR7 plays a key role in regulating SCCHN metastasis, and this process involves several molecules, such as PI3K/cdc42, pyk2, and Src. In this study, the goals are to identify whether JAK2/STAT3 also participates in CCR7's signal network, its relationship with other signal pathways, and its role in SCCHN cell invasion and migration. The results showed that stimulation of CCL19 could induce JAK2/STAT3 phosphorylation, which can be blocked by Src and pyk2 inhibitors. After activation, STAT3 was able to promote low expression of E-cadherin and had no effect on vimentin. This JAk2/STAT3 pathway not only mediated CCR7-induced cell migration but also mediated invasion speed. The immunohistochemistry results also showed that the phosphorylation of STAT3 was correlated with CCR7 expression in SCCHN, and CCR7 and STAT3 phosphorylation were all associated with lymph node metastasis. In conclusion, JAk2/STAT3 plays a key role in CCR7 regulating SCCHN metastasis.

CCR7 regulates cell migration and invasion through MAPKs in metastatic squamous cell carcinoma of head and neck.

Migration and invasion of tumor cells are essential prerequisites for the formation of metastasis in malignant diseases. Previously, we have reported that CC chemokine receptor 7 (CCR7) regulates the mobility of squamous cell carcinoma of head and neck (SCCHN) cells through several pathways, such as integrin and cdc42. In this study, we investigated the connection between CCR7 and mitogen-activated protein kinase (MAPK) family members, and their influence on cell invasion and migration in metastatic SCCHN cells. Western blotting, immunostaining and fluorescence microcopy were used to detect the protein expression and distribution of MAPKs, and the Migration assay, Matrigel invasion assay and wound-healing assay to detect the role of MAPKs in CCR7 regulating cell mobility. To analyze the correlation between CCR7 and MAPK activity and clinicopathological factors immunohistochemical staining was emplyed. The results showed stimulation of CCL19 and the activation of CCR7 could induce ERK1/2 and JNK phosphorylation, while it had no efect on p38. After activation, ERK1/2 and JNK promoted E-cadherin low expression and Vimentin high expression. The MAPK pathway not only mediated CCR7 induced cell migration, but also mediated invasion speed. The immunohistochemistry results showed that CCR7 was correlated with the phosphorylation of ERK1/2 and JNK in SCCHN, and these molecules were all associated with lymph node metastasis. Therefore, our study demonstrates that MAPK members (ERK1/2 and JNK) play a key role in CCR7 regulating SCCHN metastasis.

Submental versus platysma flap for the reconstruction of complex facial defects following resection of head and neck tumors.

OBJECTIVE: To compare the platysma flap with submental flap in terms of tumor and flap characteristics, operative properties and the functional outcomes. METHODS: A total of 65 patients presented with tumors of head and neck and underwent curative tumor resection with different neck dissections at the Department of Oromaxillofacial-Head and Neck Surgery, School of Stomatology of China Medical University; from March 2005 to December 2012 were included in the study. After radical tumor excision and neck dissection the resultant complex defects were reconstructed with either platysma flap or the submental flap. The extent of surgical resection, the type of neck dissection and choice of flap reconstruction was at the discretion of the surgical team. The functional outcomes, operative time and characteristics of both platysma and submental flaps were compared and the statistical tests of significance were applied accordingly. RESULTS: The mean age was 60 years. The complex facial defects of 30 patients were reconstructed with platysma flap and of 35 patients with submental flap. Mean operation time of submental flap including flap harvesting (5.58±1.96hrs) was shorter than platysma flap (6.2±1.4hrs). The majority of the flaps (88-93%) were taken successfully in both groups. Submental flap was associated with significantly higher patients' satisfaction regarding acceptable functional outcomes (p-value 0.027). The mean reduction in mouth opening was significantly smaller in platysma group (0.37 ±0.18cms) than the submental group (0.47±0.16). CONCLUSION: This study demonstrates that both platysma and submental flap techniques can be used for the reconstruction of complex facial defects with the acceptable functional outcome. The platysma flap can be harvested to medium size defects up to 70cm(2) with good mouth opening. The submental flap is simpler, faster with a wider range of application and more acceptable functional outcomes.

Modified visor approach applied to total or subtotal glossectomy and reconstruction: avoidance of lip splitting and mandibulotomy and cutting off mental nerve.

A lower lip-splitting incision has traditionally been performed with different types of mandibulotomy approaches for obtaining wide access to total or subtotal glossectomy. However, lip splitting can be associated with unfavorable aesthetic and function results. We describe our new modification of a traditional visor approach without lip splitting, mandibulotomy, and reserve mental nerve to avoid these morbidities and to compare aesthetic, functional, and patient subjective outcomes between the two access procedures.Of the patients undergoing total or subtotal glossectomy and reconstruction with flaps, 99 were grouped according to a surgical access procedure performed (lip split and mandibulotomy [LSM] or modified visor approach [MVA]). Data on surgical morbidity and outcomes were compared. All the tumors were safely removed by means of our modified visor approach through the combined intraoral and transcervical routes with adequate resection margins. There were no troublesome difficulties in reconstruction of the surgical defects with various flaps. Recurrence rates, swallowing, chewing, and speech were similar for both groups. Rates of postoperative fistulae were 9.3 % (LSM) vs 0 % (MVA). There were significant differences between the two groups in the temporomandibular joint (TMJ) signs (p = 0.000) and for appearance domains (p = 0.01). Avoiding lip splitting and mandibulotomy reduces patient morbidity and hospital stay and gets excellent aesthetic consequences; reserve mental nerve can avoid lower lip numbness after surgery. In our experience, the lower lip-splitting and mandibulotomy procedure for surgical exposure is unnecessary for both oncologic resection and reconstruction of tongue cancers.

Modified incision design for submental flap: an excellent design method for the reconstruction of a defect after head and neck tumor resection.

BACKGROUND: The usage of submental flap is a good method for head and neck reconstruction, but it has some risk also, such as anatomical variations and surgical errors. In this article, we present a modified incision design for the submental flap. METHODS: We designed a modified submental flap incision method based on the overlap of the incision outline of the submental flap, platysma myocutaneous flap and infrahyoid myocutaneous flap. If we found that the submental flap was unreliable during the neck dissection at the level III, II and Ib areas, the infrahyoid myocutaneous flap or platysma myocutaneous flap was used to replace it. Between 2004 and 2012, we performed 30 cases using this method. As control, 33 radial forearm free flaps were counted. Significant differences were evaluated using the χ(2) test and Mann-Whitney U. Survival and recurrence were analyzed using the Kaplan-Meier method. RESULTS: Of the 30 patients, 27 finally received a submental flap, 1 patient received an infrahyoid myocutaneous flap, and 2 patients received a platysma myocutaneous flap. In patients who received the submental flap, the average operation time was 5.9 hours, 2.4 hours shorter than the radial forearm free flap group; the average age was 61.8, 6.1 years older than the radial forearm free flap group; the survival time and recurrence time did not significantly differ with those of the forearm free flap group; and the success rate was higher than traditional methods. CONCLUSIONS: The wider indications, less required time, the similar low risk of recurrence and death as radial forearm free flap, higher success rate than traditional submental flap harvest methods, and ability to safely harvest a submental flap make the modified incision design a reliable method.

Comparison studies of different flaps for reconstruction of buccal defects.

Our purpose was to evaluate the feasibility of radial forearm free (RFF) flap, platysma myocutaneous (PM) flap, and anterolateral thigh (ALT) flap in buccal reconstruction. This study consisted of 56 patients who were categorized into 3 groups. The Student t test was used to analyze the variables. Patients in group platysma flap were significantly older, the dissection of platysma flap was easier, and the defect was significantly smaller than those in group radial forearm flap and group ALT flap. The reduction in the widths of mouth opening between group PM, group RFF, and group ALT were compared. However, the reduction of mouth-opening widths in group RFF and group ALT was significantly less than that in group PM. Platysma myocutaneous flap may be more suitable in patients with small to middle-size defect and poor status, although the flap cannot achieve a reliable result; anterolateral thigh flap and radial forearm flap can preserve the interincisal distance well even for large buccal defect, but it takes more time and skills in the operation.